柴胡疏肝散调控FXR/Nrf2/ARE通路对肝内胆汁淤积大鼠肝损伤的保护作用研究  

Study on protective effect of Chaihu Shugan Powder against liver injury in rats with intrahepatic cholestasis by regulating FXR/Nrf2/ARE pathway

作　　者：娄静[1] 赵雷[1] 朱岩洁 袁帅强 王菲[1] 张杭洲 徐娇娇 余晓珂[2] 侯留法[1] LOU Jing;ZHAO Lei;ZHU Yan-jie;YUAN Shuai-qiang;WANG Fei;ZHANG Hang-zhou;XU Jiao-jiao;YU Xiao-ke;HOU Liu-fa(Department of Liver,Gallbladder,Spleen and Stomach,Affiliated Hospital of Henan Academy of Chinese Medicine,Zhengzhou 450003,China;Department of Geriatrics,Third Affiliated Hospital of Henan University of Traditional Chinese Medicine,Zhengzhou 450003,China)

机构地区：[1]河南省中医药研究院附属医院肝胆脾胃科,河南郑州450003 [2]河南中医药大学第三附属医院老年病内科,河南郑州450003

出　　处：《中国中药杂志》2022年第20期5610-5616,共7页China Journal of Chinese Materia Medica

基　　金：河南省中医药科学研究专项(2018ZY1018,2019ZY2015,2022ZY1147);河南省中医药传承与创新人才工程(仲景工程)中医药学科领军人才项目;河南省中医药传承与创新人才工程(仲景工程)中医药学科拔尖人才项目。

摘　　要：探讨柴胡疏肝散(Chaihu Shugan Powder,CHSG)调控法尼酯衍生物X受体(farnesoid X receptor,FXR)/核因子E2相关因子(nuclear factor erythroid-2-related factor,Nrf2)/抗氧化反应元件(antioxidant response element,ARE)通路对肝内胆汁淤积大鼠肝损伤的影响。将84只SD大鼠分为正常组、模型组、CHSG-L组(0.5 g·kg^(-1))、CHSG-H组(2.5 g·kg^(-1))、熊去氧胆酸(ursodeoxycholic acid,UDCA)组(100 mg·kg^(-1))、CHSG-H+sh-NC组(2.5 g·kg^(-1)CHSG+皮下注射sh-NC慢病毒)、CHSG-H+sh-FXR组(2.5 g·kg^(-1)CHSG+皮下注射sh-FXR慢病毒),每组12只。除正常组、模型组外,其余各组均给予对应药物进行处理,每日1次,持续7 d。在第5天时除正常组外,其余组均按照100 mg·kg^(-1)的剂量灌胃α-萘异硫氰酸酯(α-naphthylisothi,ANIT),每日1次,持续3 d以构建肝内胆汁淤积大鼠模型,正常组灌胃等量生理盐水。末次给药1 h后,麻醉大鼠,进行2 h胆汁流量的测定;采用Aeroset全自动化学分析仪检测大鼠血清中谷丙转氨酶(alanine transaminase,ALT)、谷草转氨酶(aspartate aminotransferase,AST)、总胆红素(total bilirubin,TBIL)、总胆汁酸(total bile acid,TBA)的水平;HE染色检测大鼠肝组织病理变化;谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)、超氧化物歧化酶(super oxide dismutase,SOD)、丙二醛(malondialdehyde,MDA)试剂盒检测大鼠肝组织匀浆中GSH-Px、SOD、MDA水平;Western blot检测大鼠肝组织中FXR、Nrf2、血红素氧合酶1(heme oxygenase-1,HO-1)蛋白表达。与正常组比较,模型组大鼠肝组织内可见许多点状或集中的坏死区,有大量炎性细胞浸润,肝细胞明显肿胀、细胞核皱缩,2 h胆汁流量、GSH-Px、SOD水平及FXR、Nrf2、HO-1蛋白相对表达量显著降低,ALT、AST、TBIL、TBA、MDA水平显著升高;与模型组比较,CHSG-L组、CHSG-H组、UDCA组大鼠肝组织病理损伤显著减轻,2 h胆汁流量、GSH-Px、SOD水平及FXR、Nrf2、HO-1蛋白相对表达量显著升高,ALT、AST、TBThis study aims to investigate the effect of Chaihu Shugan Powder(CHSG)on liver injury in rats with intrahepatic cholestasis by regulating farnesoid X receptor(FXR)/nuclear factor erythroid-2-related factor(Nrf2)/antioxidant response element(ARE)pathway.Eighty-four SD rats were classified into normal group,model group,CHSG-L group(0.5 g·kg^(-1)),CHSG-H group(2.5 g·kg^(-1)),ursodeoxycholic acid group(UDCA group,100 mg·kg^(-1)),CHSG-H+sh-NC group(2.5 g·kg^(-1)CHSG+subcutaneous injection of sh-NC lentivirus),CHSG-H+sh-FXR group(2.5 g·kg^(-1)CHSG+subcutaneous injection of sh-FXR lentivirus),with 12 rats in each group.Rats were treated with corresponding drugs except for the normal group and the model group,once a day,for 7 days.On 5 th day,rats,except the normal group,were givenα-naphthalene isothiocyanate(ANIT)at a dose of 100 mg·kg^(-1),once a day for 3 days to induce intrahepatic cholestasis,and the normal group was given the same amount of normal saline.Rats were anesthetized 1 h after the last administration and the 2 h bile flow was measured.Aeroset chemistry analyzer was employed to detect the levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),total bilirubin(TBIL),and total bile acid(TBA)in rat serum.Based on hematoxylin and eosin(HE)staining,the pathological changes of rat liver tissue were observed.Glutathione peroxidase(GSH-Px),superoxide dismutase(SOD),and malondialdehyde(MDA)in rat liver tissue homogenate were monitored with corresponding kits.Western blot was used to detect the expression of FXR,Nrf2,and heme oxygenase-1(HO-1)proteins in rat liver tissue.Compared with the normal group,the model group showed many spots or concentrated necrotic areas in the liver tissue,infiltration of a large number of inflammatory cells,swelling liver cells with nuclear shrinkage.The 2 h bile flow,levels of GSH-Px and SOD,and relative expression of FXR,Nrf2,and HO-1 proteins were significantly lower,and the levels of ALT,AST,TBIL,TBA and MDA were significantly higher in the model group than i

关 键 词：柴胡疏肝散 肝内胆汁淤积症 氧化应激 法尼酯衍生物X受体/核因子E2相关因子/抗氧化反应元件通路 

分 类 号：R285.5[医药卫生—中药学]